CN102286376A - Microbial inoculum for high-efficiency fermenting bed and preparation method thereof - Google Patents
Microbial inoculum for high-efficiency fermenting bed and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a microbial inoculum for a high-efficiency fermenting bed and a preparation method thereof. The microbial inoculum is prepared by thoroughly and evenly mixing 30-40% of Bacillus subtilis leavening powder, 30-40% of Bacillus licheniformis leavening powder and 20-30% of Candida utilis leavening powder. The microbial inoculum can effectively promote in-situ degradation of livestock and poultry excrement, obviously reduce the generation of ammonia and other putrilages in the livestock and poultry excrement, and enhance the quality of pork.
Description
Technical field
The present invention relates to the microbial fermentation field, definite a kind of fermentation bed microbiobacterial agent and preparation method thereof of saying so.
Background technology
Along with the fast development of China's pig industry mass-producing, intensification, feces of livestock and poultry and sewage discharge are serious to showing pollution day of environment in recent years.According to statistics, the solid waste total amount that now national annual herding is produced is about 3,000,000,000 tons, only Changsha annual just has 8,000 ten thousand tons of livestock breeding wastewaters approximately, solid manure is discharged more than 1,200,000 tons, most of plant has caused bigger pollution only through just directly effluxing after the methane-generating pit simple process to environment.
The fecaluria treatment technology that present tradition is raised pigs adopts modes such as compost fermentation, biogas fermentation, aerobic fermentation drying technology to handle mostly, compost fermentation is handled the ight soil land occupation, atmospheric pollution to environment is bigger, biogas fermentation has the limitation in season, aerobic fermentation drying technology cost is higher, and certain limitation is all arranged in actual utilization.
Probiotics fermention bed to raise pig technology is according to little ecological principle and biological fermentation theory, the auxiliary starter such as agricultural crop straw, microbiobacterial agent and rice bran, sugar etc. of saw dust, rice husk, pulverizing is mixed by a certain percentage, be layered in the pig house fermentation vat, spontaneous fermentation, form the microbial fermentation bed, feeding live pig utilizes microorganism to livestock and poultry fecaluria " original position degraded " on it, reaches the novel aquaculture model of ecotope " zero pollutes ".
Because proportioning, thickness, the residing geographical ecotope difference of fermenting bed padding make that ventilation property, water-absorbent, atmospheric moisture and the envrionment temperature of bedding and padding are different; Because live pig must be degraded timely constantly producing movement, therefore select for use safety, stable, colonization ability is strong, degradation rate is high and function to have the complex micro organism fungicide that the bacterial strain of complementary action forms be the favourable assurance that fermentation bed cleaning is cultured.
Summary of the invention
Purpose of the present invention aims to provide a kind of effective promotion livestock and poultry fecaluria " original position degraded ", significantly reduces the generation of ammonia and other septic matter in the livestock and poultry fecaluria, and the fermentation bed that can improve meat quality is with microbiobacterial agent and preparation method thereof.
The objective of the invention is to realize in the following manner:
A kind of efficient fermentation bed preparation method of complex micro organism fungicide may further comprise the steps:
1) fermentation of bacillus subtilis thing preparation
At first, subtilis is cultivated through slant strains activation, shake-flask seed;
Carry out fermentor cultivation then: the substratum loading amount is 50～70% of a cumulative volume in the fermentor tank, inoculum size accounts for 0.1～0.3% of culture volume, 34～37 ℃ of leavening temperatures, blowing air 180～200rpm stir culture, the ratio of ventilation volume and fermentating liquid volume 0.9: 1～1: 1, tank pressure 0.03～0.05MPa; Fermentation termination is not less than 90% of total count for the gemma number, and the viable count of fermented liquid is 1.0 * 10
10～1.5 * 10
10Between the cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: glucose 0.5～0.7%, starch 0.3～0.5%, soybean cake powder 0.8～1.2%, manganous sulfate 0.0062%, yeast powder 0.5～0.8%, peptone 0.3～0.5%, iron(ic) chloride 0.01%, potassium primary phosphate 0.4%, lime carbonate 0.1%, sal epsom 0.05%, all the other are water, pH 7.2～7.4;
Carry out solid absorption at last: fermented liquid adsorbs back low temperature dehumidifier drying with the part by weight absorption of wheat bran by 1: 2～1: 3, pulverizes, and the powder product bacteria containing amount is 2.0 * 10
9～5.0 * 10
9Between the cfu/g;
2) the lichen bacillus ferments thing preparation
At first, Bacillus licheniformis is cultivated through slant strains activation, shake-flask seed; Described Bacillus licheniformis, preserving number are CGMCC No.4835;
Carry out fermentor cultivation then: the substratum loading amount is 50～70% of a cumulative volume in the fermentor tank, inoculum size accounts for 0.1～0.3% of culture volume, 45～50 ℃ of leavening temperatures, blowing air 180～200rpm stir culture, the ratio of ventilation volume and fermentating liquid volume 0.9: 1～1: 1, tank pressure remains on 0.03～0.05MPa, and fermentation termination is not less than 90% of total count for the gemma number, and fermented liquid contain the bacterium number 8.0 * 10
9～1.2 * 10
10Between the cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: starch 1.0～1.5%, dextrin 0.5～0.8%, soybean cake powder 1.2～1.5%, ammonium sulfate 0.6～0.8%, yeast powder 0.3～0.4%, lime carbonate 0.2%, sodium-chlor 0.4%, potassium primary phosphate 0.02%, all the other are water, pH 7.0～7.2;
Carry out solid absorption at last: fermented liquid adsorbs back low temperature dehumidifier drying with the part by weight absorption of wheat bran by 1: 2～1: 3, pulverizes, and the powder product bacteria containing amount is 1.5 * 10
9～4.0 * 10
9Between the cfu/g;
3) Candida utilis fermented product preparation
At first, Candida utilis is cultivated through slant strains activation, shake-flask seed;
Carry out fermentor cultivation then: the substratum loading amount is 50～70% of a cumulative volume in the fermentor tank, inoculum size accounts for 0.3～0.5% of culture volume, 25～28 ℃ of leavening temperatures, blowing air 120～150rpm stir culture, the ratio of ventilation volume and fermentating liquid volume 1: 1～1.1: 1, tank pressure remains on 0.03～0.05MPa, and what fermentation in 46～48 hours finished secondary fermentation liquid contains the bacterium number 2.0 * 10
9～3.0 * 10
9Between the cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: glucose 3.5～4.0%, and ammonium sulfate 1.5～2.0%, yeast powder 0.2～0.25%, potassium primary phosphate 0.1%, sodium-chlor 0.01%, sal epsom 0.05%, all the other are water, pH 6.0～6.5;
Carry out solid absorption at last: fermented liquid adsorbs back low temperature dehumidifier drying with the part by weight absorption of wheat bran by 1: 1, pulverizes, and the powder product bacteria containing amount is 1.5 * 10
9～2.5 * 10
9Between the cfu/g;
4) efficient fermentation bed is composite with complex micro organism fungicide
Careless by weight percentage fermentation of bacillus medicinal powder 30～40%, the lichen bacillus ferments medicinal powder 30～40%, Candida utilis fermented product pulvis 20%～30%, fully mixing gets final product.
Described efficient fermentation bed is not less than 1.0 * 10 with the total viable count of complex micro organism fungicide
9Cfu/g.
Each constituent mass per-cent of the described slant strains activatory of step 1) substratum is: peptone 1%, and NaCl 0.5%, yeast extract paste 0.5%, agar 1.5%～2.0%, all the other are water, pH 7.2-7.4;
The substratum that the described shake-flask seed of step 1) is cultivated is not add agar in the described slant medium of step 1), concrete operations: get fresh slant strains one ring of activatory, be inoculated in the shake-flask seed liquid substratum, loading amount is 100mL/500mL, rotating speed is 180～200rpm, behind 34～37 ℃ of constant temperature culture 22～24h, 80 ℃ of water-bath thermal treatment 10 minutes gets final product.
Step 2) described each constituent mass per-cent of slant strains activatory substratum is: glucose 0.4%, and malt extract 1%, yeast extract 0.4%, lime carbonate 0.2%, agar 1.5%～2.0%, all the other are water, pH 7.0～7.2;
Step 2) substratum cultivated of described shake-flask seed is in step 2) do not add agar in the described slant medium, concrete operations: get fresh slant strains one ring of activatory, be inoculated in the shake-flask seed liquid substratum, loading amount is 100mL/500mL, rotating speed is 180～200rpm, behind 45～50 ℃ of constant temperature culture 28～30h, stop cultivating.
The described slant strains activatory of step 3) substratum is the PDA substratum;
The described shake-flask seed of step 3) is cultivated concrete operations: get fresh slant strains one ring of activatory, be inoculated in the shake-flask seed liquid substratum, loading amount is 150mL/500mL, and rotating speed is 120～150rpm, behind 25～28 ℃ of constant temperature culture 30～32h, stops cultivating; Each constituent mass per-cent of the substratum of described shake-flask seed: glucose 2%, peptone 2%, yeast extract 1%, all the other are water, pH 6.0～6.5.
A kind of efficient fermentation bed is the microbial inoculum that is prepared from by above-mentioned method with microbiobacterial agent.Described Bacillus licheniformis, preserving number are CGMCC No.4835.
Advantage of the present invention:
The selection of excellent species
Subtilis: aerobic bacteria, can produce proteolytic enzyme, multiple enzyme such as amylase can obviously improve growth of animal speed and efficiency of feed utilization.In the feed course of processing and sour environment advantages of higher stability is arranged, reproduction speed is fast, good stress resistance, and decomposition fecaluria ability is strong.Through the bacterial strain optimal temperature after optimizing is 30 ℃～37 ℃, is inoculated in the liquid fermentation medium, produces proteolytic enzyme during 24h and produce amylase activity to be respectively 2706u/mL, 31.331MMU; Be seeded in the pig manure urine extracting solution and react after 25 days, degradation rate, the NH of COD
3-N degradation rate is respectively 86.5%, 70.7%.
Bacillus licheniformis: screening and separating obtains the high growth temperature bacterium from the pig manure that becomes thoroughly decomposed of health pig discharging.Can produce proteolytic enzyme, amylase, cellulase etc.At hot stage, bacterium is more active.Through the bacterial strain optimal temperature after optimizing is 45 ℃～55 ℃, is seeded in the pig manure urine extracting solution to react after 25 days, to NH
3-N degradation rate is 75.2%.
Candida utilis: the oxygen bacterium of holding concurrently, 25 ℃～28 ℃ of suitable growth temperatures, pH value 6.0～6.5.For animal provides protein and VITAMIN, help digest, stimulate growth of animal, suppress the pathogeny microbial reproduction, enhance immunity power.Be seeded in the pig manure urine extracting solution and react after 2 days, NH
3-N degradation rate is 83.5%.
Product of the present invention fermentation bed with complex micro organism fungicide be seeded in the pig manure urine extracting solution reaction after 25 days the COD degradation rate reached 89.9%, NH
3-N clearance reaches 84.5%.
The preservation information of bacterial strain of the present invention is as follows:
Preservation date: on May 9th, 2011
Depositary institution's title: China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC)
Deposit number: CGMCC No.4835
Classification name: Bacillus licheniformis (Bacillus licheniformis).
Beneficial effect of the present invention
Product of the present invention is according to the having complementary functions property structure of the useful bacterial strain of difference, and it forms clear and definite, steady quality.After particularly passing through the unique zymotechnique of the present invention, what in fact obtain is the mixture of microbial inoculum and fermentating metabolism product, may can produce many useful materials.Production testing by reality finds that product of the present invention is used for fermentation bed to raise pig and can utilizes microorganism to livestock and poultry fecaluria " original position degraded ", reaches to reduce the pollution of pig farm to ecotope, realizes the recycling ecological agriculture purpose; Simultaneously can improve the live pig welfare, improve meat quality.
Embodiment
Following embodiment is intended to further specify the present invention, and unrestricted the present invention.
3 strain strain name that the present invention adopts and bacterial classification source are as follows:
Subtilis (ACCC10619) is bought in Chinese agriculture microbial strains preservation center;
Bacillus licheniformis (Bacillus licheniformis), screening and separating obtains from the pig manure that becomes thoroughly decomposed of health pig discharging, identifies through institute of microbiology of the Chinese Academy of Sciences.And carried out preservation at China Committee for Culture Collection of Microorganisms common micro-organisms center on May 9th, 2011, and Bacillus licheniformis (Bacillus licheniformis), preserving number is CGMCCNo.4835.
Screening process is as follows: healthy pig manure sample that will become thoroughly decomposed and enrichment medium (fresh pig manure) be (weight ratio) thorough mixing by a certain percentage, progressively add wood sawdust and rice husk, the water ratio of regulating material is about 55%, be loaded in the clean plastic tank, static incubated at room temperature, stir every day for several times, measure foul smell variation and weight-loss ratio changing conditions in the culturing process, when the basic odorless of fecaluria and loss of weight are not obvious, culture transferring continues shaking culture, progressively improve enrichment medium concentration to 95%, each is taken turns and is cultured to odorless, when loss of weight is not obvious till.The enrichment culture thing by repeatedly dull and stereotyped coating separation, is obtained strong bacterial strain 4 strains of fecaluria capacity of decomposition.Wherein 1 strain bacterial strain bacterium colony is bigger, and the edge is irregular; It is shaft-like that thalline is, and gemma is arranged, Gram-positive; Starch hydrolysis experiment, gelatin hydrolysis, oxidase test, catalase test, methyl red test, citrate test, VP＜pH6 test positive; VP＞pH7 tests negative.Be accredited as Bacillus licheniformis through institute of microbiology of the Chinese Academy of Sciences.
Candida utilis (ACCC20060) is bought in Chinese agriculture microbial strains preservation center.
Embodiment 1: the composite fungus agent preparation
1. fermentation of bacillus subtilis thing preparation
The slant strains activation: get refrigerator and preserve slant strains, aseptic technique is inoculated in the fresh test tube slant substratum, and 37 ℃ of constant temperature culture are spent the night, and make fresh test tube slant bacterial classification.Slant medium: peptone 1%, NaCl 0.5%, yeast extract paste 0.5%, agar 1.5%～2.0%, all the other are water, pH 7.2-7.4.
Shake-flask seed is cultivated: get fresh slant strains one ring of activatory, aseptic technique is inoculated in the shake-flask seed liquid substratum, and loading amount is 100mL/500mL, rotating speed is 180～200rpm, behind 34～37 ℃ of constant temperature culture 22～24h, 80 ℃ of water-bath thermal treatment 10 minutes gets final product.The substratum of described shake-flask seed is not add agar in slant medium.
Fermentor cultivation: the substratum loading amount is 50～70% of a cumulative volume in the fermentor tank, inoculum size accounts for 0.1～0.3% of culture volume, 34～37 ℃ of leavening temperatures, blowing air stir culture (180～200rpm), ventilation volume was than 0.9: 1～1: 1, tank pressure remains on 0.03-0.05MPa, and free air delivery and tank pressure are suitably adjusted according to the foam situation in the inoculation back, prevent to escape liquid.After foam fades away, strengthen air flow.Per 4 hours sampling microscopies once.Fermentation termination accounts for more than 90% of total count for the gemma number.Put a jar sample and measure total count with colony counting method.Fermented liquid contain the bacterium number 1.0 * 10
10-1.5 * 10
10Between the cfu/ml.Fermentation tank culture medium: glucose 0.5～0.7%, starch 0.3～0.5%, soybean cake powder 0.8～1.2%, manganous sulfate 0.0062%, yeast powder 0.5～0.8%, peptone 0.3～0.5%, iron(ic) chloride 0.01%, potassium primary phosphate 0.4%, lime carbonate 0.1%, sal epsom 0.05%, all the other are water, pH 7.2～7.4.
Solid absorption: the part by weight that fermented liquid was pressed 1: 2～1: 3 with wheat bran adsorbs, and low temperature dehumidifier drying in absorption back is pulverized, and behind the mensuration bacteria containing amount, puts in storage and preserves.The powder product bacteria containing amount is 2.0 * 10
9-5.0 * 10
9Between the cfu/g.
2. the lichen bacillus ferments thing preparation
Each constituent mass per-cent of slant strains activatory substratum is: glucose 0.4%, and malt extract 1%, yeast extract 0.4%, lime carbonate 0.2%, agar 1.5%～2.0%, all the other are water, pH 7.0～7.2;
Get fresh slant strains one ring of activatory, be inoculated in the shake-flask seed liquid substratum, loading amount is 100mL/500mL, and rotating speed is 180～200rpm, behind 45～50 ℃ of constant temperature culture 28～30h, stops cultivating.The substratum of described shake-flask seed is not add agar in slant medium.
Carry out fermentor cultivation then: the substratum loading amount is 50～70% of a cumulative volume in the fermentor tank, inoculum size accounts for 0.1～0.3% of culture volume, 45～50 ℃ of leavening temperatures, blowing air stir culture (180～200rpm), ventilation volume was than (ratio of air flow and fermentating liquid volume) 0.9: 1～1: 1, tank pressure remains on 0.03～0.05MPa, and fermentation termination is not less than 90% of total count for the gemma number; Fermented liquid contain the bacterium number 8.0 * 10
9～1.2 * 10
10Between the cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: starch 1.0～1.5%, dextrin 0.5～0.8%, soybean cake powder 1.2～1.5%, ammonium sulfate 0.6～0.8%, yeast powder 0.3～0.4%, lime carbonate 0.2%, sodium-chlor 0.4%, potassium primary phosphate 0.02%, all the other are water, pH 7.0～7.2;
Carry out solid absorption at last: fermented liquid adsorbs back low temperature dehumidifier drying with the part by weight absorption of wheat bran by 1: 2～1: 3, pulverizes, and the powder product bacteria containing amount is 1.5 * 10
9～4.0 * 10
9Between the cfu/g.
3. Candida utilis fermented product preparation
Slant strains activatory substratum is the PDA substratum;
Get fresh slant strains one ring of activatory, be inoculated in the shake-flask seed liquid substratum, loading amount is 150mL/500mL, and rotating speed is 120～150rpm, behind 25～28 ℃ of constant temperature culture 30～32h, stops cultivating.Each constituent mass per-cent of the substratum of described shake-flask seed: glucose 2%, peptone 2%, yeast extract 1%, all the other are water, pH 6.0～6.5.
Carry out fermentor cultivation then: the substratum loading amount is 50～70% of a cumulative volume in the fermentor tank, inoculum size accounts for 0.3～0.5% of culture volume, 25～28 ℃ of leavening temperatures, blowing air stir culture (120～150rpm), ventilation volume was than (ratio of air flow and fermentating liquid volume) 1: 1～1.1: 1, tank pressure remains on 0.03～0.05MPa, and fermentation in 46～48 hours finishes; Fermented liquid contain the bacterium number 2.0 * 10
9～3.0 * 10
9Between the cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: glucose 3.5～4.0%, and ammonium sulfate 1.5～2.0%, yeast powder 0.2～0.25%, potassium primary phosphate 0.1%, sodium-chlor 0.01%, sal epsom 0.05%, all the other are water, pH 6.0～6.5;
Carry out solid absorption at last: fermented liquid adsorbs back low temperature dehumidifier drying with the part by weight absorption of wheat bran by 1: 1, pulverizes, and the powder product bacteria containing amount is 1.5 * 10
9～2.5 * 10
9Between the cfu/g.
4. zymophyte is composite
The fermentation of bacillus subtilis medicinal powder 30～40% by weight percentage, the lichen bacillus ferments medicinal powder 30～40%, and Candida utilis fermented product pulvis 20～30%, fully mixing final vacuum packing promptly obtains composite fungus agent.
Embodiment 2: the effect of raising pigs of utilizing composite fungus agent preparation fermentation bed of the present invention
1. fermentation bed preparation
Fermenting bed padding is made of by 1: 1 weight ratio saw dust, rice husk, or is made of by 5: 3: 2 weight ratio saw dust, husk, agricultural crop straw.Saw dust, husk, agricultural crop straw play the absorption microbial inoculum, keep moisture, loosen and breathe freely, provide for functional microorganism effects such as C, N source.Preferably saw dust, rice husk, stalk etc. were tanned by the sun under the sun 1～2 day before making the fermentation bed, the hurdle house is wanted thorough disinfection.
Fermenting bed padding should not have go mouldy, do not have corrupt, ordorless or other peculiar smell.Material particular diameter is at 2～10mm, carbon nitrogen ratio 40～100: 1, and colibacillus of excrement number≤100cfu/g, induced worm egg death rate 〉=98%, pH value about 7.5.Chemical glue plywood saw dust can not be used as bedding and padding.
In fermentation vat, product of the present invention, bedding and padding, auxiliary starter are added water and mix, be layered in the fermentation vat, the moisture content about 45% of bedding and padding, about heap fermentation 3 days, when bedding and padding middle level temperature reaches 45 ℃～55 ℃, plough deeply once, ferment again about 3 days, when treating that the middle level temperature is reduced to 40 ℃ of left and right sides, the bedding and padding of piling up are raked, meet the requirements and just can advance pig, undesirable then fermentation sterilization again.Thickness is 80cm after the fermenting bed padding compacting, than about the low 10cm of the table top of searching for food.
2. test grouping
Carried out product fermentation bed cultivation technology pig-keeping experiment of the present invention 5 days～December 8 July in 2009 on certain pig farm.Bedding and padding are made of by 1: 1 weight ratio saw dust, rice husk.By term, the principle that body weight is close, select same kind, with batch, 180 of hybridization (the local white ♀ * big York ♂) pigs of mean body weight (16.35 ± 2.33) kg, every pig hangs up ear tag, is divided into 3 groups at random, every group of 4 repetition, 8 boars of every repetition, 7 sows.Test I group is control group, traditional water mud ground hutch; Test II group, the fermentation bed cultivation made from complex micro organism fungicide of the present invention, microbial inoculum component and weight percent thereof consist of: fermentation of bacillus subtilis medicinal powder 35%, the lichen bacillus ferments medicinal powder 35%, Candida utilis fermented product pulvis 30%, the consumption 200g/ cubic meter bedding and padding of starter; Test III group, the fermentation bed cultivation made from complex micro organism fungicide of the present invention, microbial inoculum component and weight percent thereof consist of: fermentation of bacillus subtilis medicinal powder 40%, the lichen bacillus ferments medicinal powder 40%, Candida utilis fermented product pulvis 20%, the consumption 200g/ cubic meter bedding and padding of starter.
3. experimental result is as follows:
Table 1 colony house environment and live pig surface observation result
Table 2 live pig production performance result
By table 1, table 2 as can be seen, the colony house environment of test II, III group is obviously compared according to I and is organized, and does not have obvious stink, does not have obvious mosquitos and flies and grows; The average daily gain of test II, III group is higher than contrast I group (P＜0.05), and feedstuff-meat ratio is lower than control group (P＜0.05), and difference is not remarkable between test II, the III group.Reach a conclusion thus, microbial inoculum of the present invention can alleviate environmental pollution, improves efficiency of feed utilization, effectively promotes the growth of pig.
Table 3 raising pattern is to the influence of live pig meat
As can be seen from Table 3, the test group meat quality is better than control group.Simultaneously pork is checked through livestock product quality safety supervision and inspection center of the Ministry of Agriculture (Changsha), each test group pork meat meets " no public nuisance livestock meat safety requirements standard (GB18406.3-2001) " requirement fully, illustrates that fermentation bed to raise pig has no adverse effects to pork quality.
Table 4 bedding and padding temperature variation
As can be seen from Table 4,5cm place under test II, the III group bedding and padding top layer, mean daily temperature is respectively 28.5 ℃, 29.0 ℃, and difference is not remarkable; The mean daily temperature at 30cm place, test II group is 40.9 ℃, and test III group is 44.3 ℃, and the bedding and padding nexine keeps higher temperature, helps the rapid degraded of pig manure urine.
Useful viable count changes in table 5 bedding and padding
As can be seen from Table 5, when fermentation entered 30d, it was 10 that test II group is counted content with III group probiotics
8The order of magnitude; During fermentation 150d, test II group is counted content with III group probiotics and is still kept 10
7The order of magnitude illustrates that product of the present invention has stronger field planting ability in fermenting bed padding.
Claims (7)
1. an efficient fermentation preparation method with complex micro organism fungicide is characterized in that, may further comprise the steps:
1) fermentation of bacillus subtilis thing preparation
At first, subtilis is cultivated through slant strains activation, shake-flask seed;
Carry out fermentor cultivation then: the substratum loading amount is 50～70% of a cumulative volume in the fermentor tank, inoculum size accounts for 0.1～0.3% of culture volume, 34～37 ℃ of leavening temperatures, blowing air 180～200rpm stir culture, the ratio of ventilation volume and fermentating liquid volume 0.9: 1～1: 1, tank pressure 0.03～0.05MPa; Fermentation termination is not less than 90% of total count for the gemma number, and the viable count of fermented liquid is 1.0 * 10
10～1.5 * 10
10Between the cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: glucose 0.5～0.7%, starch 0.3～0.5%, soybean cake powder 0.8～1.2%, manganous sulfate 0.0062%, yeast powder 0.5～0.8%, peptone 0.3～0.5%, iron(ic) chloride 0.01%, potassium primary phosphate 0.4%, lime carbonate 0.1%, sal epsom 0.05%, all the other are water, pH 7.2～7.4;
Carry out solid absorption at last: fermented liquid adsorbs back low temperature dehumidifier drying with the part by weight absorption of wheat bran by 1: 2～1: 3, pulverizes, and the powder product bacteria containing amount is 2.0 * 10
9～5.0 * 10
9Between the cfu/g;
2) the lichen bacillus ferments thing preparation
At first, Bacillus licheniformis is cultivated through slant strains activation, shake-flask seed; Described Bacillus licheniformis, preserving number are CGMCC No.4835;
Carry out fermentor cultivation then: the substratum loading amount is 50～70% of a cumulative volume in the fermentor tank, inoculum size accounts for 0.1～0.3% of culture volume, 45～50 ℃ of leavening temperatures, blowing air 180～200rpm stir culture, the ratio of ventilation volume and fermentating liquid volume 0.9: 1～1: 1, tank pressure remains on 0.03～0.05MPa, and fermentation termination is not less than 90% of total count for the gemma number, and fermented liquid contain the bacterium number 8.0 * 10
9～1.2 * 10
10Between the cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: starch 1.0～1.5%, dextrin 0.5～0.8%, soybean cake powder 1.2～1.5%, ammonium sulfate 0.6～0.8%, yeast powder 0.3～0.4%, lime carbonate 0.2%, sodium-chlor 0.4%, potassium primary phosphate 0.02%, all the other are water, pH 7.0～7.2;
Carry out solid absorption at last: fermented liquid adsorbs back low temperature dehumidifier drying with the part by weight absorption of wheat bran by 1: 2～1: 3, pulverizes, and the powder product bacteria containing amount is 1.5 * 10
9～4.0 * 10
9Between the cfu/g;
3) Candida utilis fermented product preparation
At first, Candida utilis is cultivated through slant strains activation, shake-flask seed;
Carry out fermentor cultivation then: the substratum loading amount is 50～70% of a cumulative volume in the fermentor tank, inoculum size accounts for 0.3～0.5% of culture volume, 25～28 ℃ of leavening temperatures, blowing air 120～150rpm stir culture, the ratio of ventilation volume and fermentating liquid volume 1: 1～1.1: 1, tank pressure remains on 0.03～0.05MPa, and what fermentation in 46～48 hours finished secondary fermentation liquid contains the bacterium number 2.0 * 10
9～3.0 * 10
9Between the cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: glucose 3.5～4.0%, and ammonium sulfate 1.5～2.0%, yeast powder 0.2～0.25%, potassium primary phosphate 0.1%, sodium-chlor 0.01%, sal epsom 0.05%, all the other are water, pH 6.0～6.5;
Carry out solid absorption at last: fermented liquid adsorbs back low temperature dehumidifier drying with the part by weight absorption of wheat bran by 1: 1, pulverizes, and the powder product bacteria containing amount is 1.5 * 10
9～2.5 * 10
9Between the cfu/g;
4) efficient fermentation bed is composite with complex micro organism fungicide
Careless by weight percentage fermentation of bacillus medicinal powder 30～40%, the lichen bacillus ferments medicinal powder 30～40%, Candida utilis fermented product pulvis 20%～30%, fully mixing gets final product.
2. method according to claim 1 is characterized in that, described efficient fermentation bed is not less than 1.0 * 10 with the total viable count of complex micro organism fungicide
9Cfu/g.
3. method according to claim 1 is characterized in that,
Each constituent mass per-cent of the described slant strains activatory of step 1) substratum is: peptone 1%, and NaCl 0.5%, yeast extract paste 0.5%, agar 1.5%～2.0%, all the other are water, pH 7.2-7.4;
The substratum that the described shake-flask seed of step 1) is cultivated is not add agar in the described slant medium of step 1), concrete operations: get fresh slant strains one ring of activatory, be inoculated in the shake-flask seed liquid substratum, loading amount is 100mL/500mL, rotating speed is 180～200rpm, behind 34～37 ℃ of constant temperature culture 22～24h, 80 ℃ of water-bath thermal treatment 10 minutes gets final product.
4. method according to claim 1 is characterized in that,
Step 2) described each constituent mass per-cent of slant strains activatory substratum is: glucose 0.4%, and malt extract 1%, yeast extract 0.4%, lime carbonate 0.2%, agar 1.5%～2.0%, all the other are water, pH 7.0～7.2;
Step 2) substratum cultivated of described shake-flask seed is in step 2) do not add agar in the described slant medium, concrete operations: get fresh slant strains one ring of activatory, be inoculated in the shake-flask seed liquid substratum, loading amount is 100mL/500mL, rotating speed is 180～200rpm, behind 45～50 ℃ of constant temperature culture 28～30h, stop cultivating.
5. method according to claim 1 is characterized in that,
The described slant strains activatory of step 3) substratum is the PDA substratum;
The described shake-flask seed of step 3) is cultivated concrete operations: get fresh slant strains one ring of activatory, be inoculated in the shake-flask seed liquid substratum, loading amount is 150mL/500mL, and rotating speed is 120～150rpm, behind 25～28 ℃ of constant temperature culture 30～32h, stops cultivating; Each constituent mass per-cent of the substratum of described shake-flask seed: glucose 2%, peptone 2%, yeast extract 1%, all the other are water, pH 6.0～6.5.
6. an efficient fermentation bed is used microbiobacterial agent, it is characterized in that, and be by any microbial inoculum that described method is prepared from of claim 1-5.
7. a strain is used for efficient fermentation bed with microbiobacterial agent fermentative preparation bacterial strain, it is characterized in that described bacterial strain is Bacillus licheniformis (Bacillus licheniformis), and preserving number is CGMCC No.4835.
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